We developed a simple and efficient synthesis for 4-pyridoxolactone starting with pyridoxine and using a whole-cell biotransformation by two transformed Escherichia coli cell types. One set of transformed cells expressed pyridoxine 4-oxidase, catalase, and chaperonin, while the second set expressed pyridoxal 4-dehydrogenase. With this combination of cells, pyridoxine was first oxidized to pyridoxal, which was then dehydrogenated to 4-pyridoxolactone by pyridoxine 4-oxidase and pyridoxal 4-dehydrogenase, respectively. In a reaction mixture containing the two transformed cell types, 10 mM of pyridoxine was completely converted into 4-pyridoxolactone at 30 degrees C in 24 h. When starting with 80 mM of pyridoxine, it was necessary to add 0.5 mM or more of NAD(+) to complete the reaction.
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http://dx.doi.org/10.1263/jbb.106.460 | DOI Listing |
Acta Crystallogr F Struct Biol Commun
April 2014
Laboratory of Applied Structural Biology, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Gokasyo, Uji, Kyoto 611-0011, Japan.
4-Pyridoxolactonase from Mesorhizobium loti catalyzes the zinc-dependent lactone-ring hydrolysis of 4-pyridoxolactone (4PAL) to 4-pyridoxic acid (4PA) in vitamin B6 degradation pathway I. The crystal structures of 4-pyridoxolactonase and its complex with 5-pyridoxolactone (5PAL; the competitive inhibitor) were determined. The overall structure was an αβ/βα sandwich fold, and two zinc ions were coordinated.
View Article and Find Full Text PDFBiosci Biotechnol Biochem
June 2011
Faculty of Agriculture and Agricultural Science Program, Graduate School of Integral Arts and Science, Kochi University, Nankoku, Kochi, Japan.
Pyridoxal 4-dehydrogenase catalyzes the irreversible oxidation of pyridoxal to 4-pyridoxolactone and is involved in degradation pathway I of pyridoxine, a vitamin B(6) compound. Its crystal structure was elucidated for the first time. Molecular replacement with (S)-1-phenylthanol dehydrogenase (PDB code 2EW8) was adopted to determine the tertiary structure of the NAD(+)-bound enzyme.
View Article and Find Full Text PDFJ Nutr Sci Vitaminol (Tokyo)
November 2010
Faculty of Agriculture, Graduate School of Integrated Arts and Sciences, Kochi University, Nankoku, Kochi, Japan.
A method for determining all of the six natural vitamin B(6) compounds and pyridoxine-beta-glucoside in urine from humans consuming their usual diet was developed. These compounds were specifically converted with 5 enzymes into a high fluorescent 4-pyridoxolactone which was supersensitively determined by an isocratic HPLC. All of the compounds in urine from humans consuming their usual diets were for the first time determined together.
View Article and Find Full Text PDFJ Biosci Bioeng
November 2008
Department of Agriculture, Faculty of Agriculture, Kochi University, 200 Monobe-Otsu, Nankoku, Kochi, Japan.
We developed a simple and efficient synthesis for 4-pyridoxolactone starting with pyridoxine and using a whole-cell biotransformation by two transformed Escherichia coli cell types. One set of transformed cells expressed pyridoxine 4-oxidase, catalase, and chaperonin, while the second set expressed pyridoxal 4-dehydrogenase. With this combination of cells, pyridoxine was first oxidized to pyridoxal, which was then dehydrogenated to 4-pyridoxolactone by pyridoxine 4-oxidase and pyridoxal 4-dehydrogenase, respectively.
View Article and Find Full Text PDFJ Nutr Sci Vitaminol (Tokyo)
February 2008
Department of Bioresources Science, Faculty of Agriculture, Kochi University, Kochi, Japan.
A determination method for individual natural vitamin B(6) compounds was developed. The vitamin B(6) compounds were specifically converted into 4-pyridoxolactone (PAL), a highly fluorescent compound, through a combination of enzymatic reactions and HCl-hydrolysis. PAL was then determined by HPLC.
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